Dual-acting stapled peptides target both HIV-1 entry and assembly

Zhang, H. et al. (2013) Dual-acting stapled peptides target both HIV-1 entry and assembly. Retrovirology, 10(136), (doi: 10.1186/1742-4690-10-136)

Abstract

Background: Previously, we reported the conversion of the 12-mer linear and cell-impermeable peptide CAI to a cell-penetrating peptide NYAD-1 by using an i,i + 4 hydrocarbon stapling technique and confirmed its binding to the C-terminal domain (CTD) of the HIV-1 capsid (CA) protein with an improved affinity (Kd ~ 1 μM) compared to CAI (Kd ~ 15 μM). NYAD-1 disrupts the formation of both immature- and mature-like virus particles in in vitro and cell-based assembly assays. In addition, it displays potent anti-HIV-1 activity in cell culture against a range of laboratory-adapted and primary HIV-1 isolates.<p></p> Results: In this report, we expanded the study to i,i + 7 hydrocarbon-stapled peptides to delineate their mechanism of action and antiviral activity. We identified three potent inhibitors, NYAD-36, -66 and -67, which showed strong binding to CA in NMR and isothermal titration calorimetry (ITC) studies and disrupted the formation of mature-like particles. They showed typical α-helical structures and penetrated cells; however, the cell penetration was not as efficient as observed with the i,i + 4 peptides. Unlike NYAD-1, the i,i + 7 peptides did not have any effect on virus release; however, they impaired Gag precursor processing. HIV-1 particles produced in the presence of these peptides displayed impaired infectivity. Consistent with an effect on virus entry, selection for viral resistance led to the emergence of two mutations in the gp120 subunit of the viral envelope (Env) glycoprotein, V120Q and A327P, located in the conserved region 1 (C1) and the base of the V3 loop, respectively.<p></p> Conclusion: The i,i + 7 stapled peptides derived from CAI unexpectedly target both CA and the V3 loop of gp120. This dual-targeted activity is dependent on their ability to penetrate cells as well as their net charge. This mechanistic revelation will be useful in further modifying these peptides as potent anti-HIV-1 agents.<p></p>

• Item Type: Articles
• Keywords: HIV-1; Capsid; Virus assembly; Virus entry; Stapled peptides; NMR; SPR; ITC; Drug-resistance
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Cooper, Professor Alan
• Authors: Zhang, H., Curreli, F., Waheed, A.A., Mercredi, P.Y., Mehta, M., Bhargava, P., Scacalossi, D., Tong, X., Lee, S., Cooper, A., Summers, M.F., Freed, E.O., and Debnath, A.K.
• Subjects: Q Science > QD Chemistry; Q Science > QH Natural history > QH345 Biochemistry; Q Science > QR Microbiology > QR355 Virology; R Medicine > R Medicine (General)
• College/School: College of Science and Engineering > School of Chemistry
• Journal Name: Retrovirology
• Publisher: BioMed Central
• ISSN: 1742-4690
• ISSN (Online): 1742-4690
• Copyright Holders: Copyright © 2013 The Authors
• First Published: First published in Retrovirology 10(136)
• Publisher Policy: Reproduced under a Creative Commons License